LOCUS       sequence-319367-        8352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses KCNMB4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8352)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: IDG_KCNMB4_OE_1
FEATURES             Location/Qualifiers
     source          1..8352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        30..333
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        334..537
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     488..508
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    560..584
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1215..1239)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1241..1282
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1324..1912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1377..1397)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1795..1806)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1984..2217
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2295..2416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2332..2351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2386..2405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2456..2591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2611..2630)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2612..2630)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2640..2657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2640..2656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2649..2671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2798..3253
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2885..2904)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3095..3116
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3279..3383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3384..3452,3453..4244)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3602..3621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4415..5003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4904..4923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5157..5174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5291..5312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5327..5332,5333..5350,5351..5357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5365..5381
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5370..5392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5389..5405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5389..5405
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5424..5444
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5426..5444
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5472..5698
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5699..5879
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5926..6051
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6544..6777
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6962..7006
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7181..7681
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7600..7618
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7693..8088
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8148..8265
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
       61 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      121 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      181 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      241 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      301 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      361 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      421 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      481 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      541 tggctaactg tcgggatcaa caagtttgta caaaaaagtt ggcatggcga agctccgggt
      601 ggcttacgag tacacggaag ccgaggacaa gagcatccgg ctcggcttgt ttctcatcat
      661 ctccggcgtc gtgtcgctct tcatcttcgg cttctgctgg ctgagtcccg cgctgcagga
      721 tctgcaagcc acggaggcca attgcacggt gctgtcggtg cagcagatcg gcgaggtgtt
      781 cgagtgcacc ttcacctgtg gcgccgactg caggggcacc tcgcagtacc cctgcgtcca
      841 ggtctacgtg aacaactctg agtccaactc tagggcgctg ctgcacagcg acgagcacca
      901 gctcctgacc aaccccaagt gctcctatat ccctccctgt aagagagaaa atcagaagaa
      961 tttggaaagt gtcatgaatt ggcaacagta ctggaaagat gagattggtt cccagccatt
     1021 tacttgctat tttaatcaac atcaaagacc agatgatgtg cttctgcatc gcactcatga
     1081 tgagattgtc ctcctgcatt gcttcctctg gcccctggtg acatttgtgg tgggcgttct
     1141 cattgtggtc ctgaccatct gtgccaagag cttggcggtc aaggcggaag ccatgaagaa
     1201 gcgcaagttc tcttacccaa ctttcttgta caaagtggtt ggtaagccta tccctaaccc
     1261 tctcctcggt ctcgattcta cgtagtaatg agctagcgct aaccggtggc gcgttaagtc
     1321 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1381 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1441 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1501 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1561 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1621 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1681 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     1741 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1801 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1861 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     1921 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     1981 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     2041 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     2101 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     2161 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     2221 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     2281 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2341 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2401 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     2461 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2521 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2581 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     2641 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2701 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2761 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     2821 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2881 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2941 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3001 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3061 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3121 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3181 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3241 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3301 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3361 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3421 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3481 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3541 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3601 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3661 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3721 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3781 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3841 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3901 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3961 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4021 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4081 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4141 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4201 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4261 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4321 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4381 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4441 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4501 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4561 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4621 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4681 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4741 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4801 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4861 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4921 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4981 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5041 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5101 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5161 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5221 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5281 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5341 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5401 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     5461 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     5521 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     5581 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     5641 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     5701 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     5761 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     5821 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     5881 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     5941 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     6001 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     6061 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     6121 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     6181 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     6241 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     6301 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     6361 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     6421 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     6481 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     6541 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     6601 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     6661 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     6721 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     6781 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     6841 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     6901 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     6961 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     7021 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     7081 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     7141 attatcgttt cagacccacc tcccaacccc gaggggaccc ttgcgccttt tccaaggcag
     7201 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     7261 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     7321 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     7381 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     7441 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     7501 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     7561 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     7621 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     7681 gggggtacca ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
     7741 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
     7801 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
     7861 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
     7921 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
     7981 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     8041 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcct gcagctgcag
     8101 tagtaagaat tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa
     8161 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     8221 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     8281 gacagcagag atccactttg gcgccggctc gagggggccc ggggttatta atagtaatca
     8341 attacggggt ca
//
